Thermal stability of apolipoprotein B100 in low-density lipoprotein is disrupted at early stages of oxidation while neutral lipid core organization is conserved.
The time course of the unfolding characteristics of the protein moiety and of the thermotropic behavior of the core-located apolar lipids of highly homogeneous low-density lipoprotein (LDL) subspecies (d 1.030-1.040 g/mL) have been evaluated during transition metal- and azo radical-induced oxidation using differential scanning calorimetry. Apolipoprotein B100 (apo-B100) structure was highly sensitive to oxidative modification; indeed, a significant loss of thermal stability was observed at initial stages irrespective of whether oxidation was mediated by site-specific binding of copper ions or by free radicals generated during decomposition of azo compounds. Subsequently, thermal protein integrity was destroyed, as a result of potentially irreversible protein unfolding, cross-linking reactions, and aggregation. Our results suggest that even minimal oxidative modification of apo-B100 has a major impact on the stability of this large monomeric protein. By contrast, the core lipids, which consist primarily of cholesteryl esters and triglycerides and play a determinant role in the thermal transition occurring near physiological temperature, preserved features of an ordered arrangement even during propagation of lipid peroxidation.
Prassl
R
R
Institute of Biophysics, Austrian Academy of Sciences, Graz.
Schuster
B
B
Laggner
P
P
Flamant
C
C
Nigon
F
F
Chapman
M J
MJ
D000578:Amidines / Q000494:pharmacology; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000737:chemistry / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning; D003300:Copper / Q000494:pharmacology; D006358:Hot Temperature; D006801:Humans; D050356:Lipid Metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008055:Lipids / Q000737:chemistry; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D000578:Amidines; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D008055:Lipids; D008077:Lipoproteins, LDL; C046728:2,2'-azobis(2-amidinopropane); D003300:Copper